MicroRNA-199b Deregulation Shows Oncogenic Properties and Promising Clinical Value as Circulating Marker in Locally Advanced Rectal Cancer Patients
Abstract
:1. Introduction
2. Results
2.1. MiR-199b Regulates Cell Migration and Agressiveness
2.2. MiR-199b Induces Apoptosis and Inhibts Colonosphere Formation
2.3. Analysis of miR-199b Expression Levels as Circulating Markers in Larc Patients and Its Association with Molecular and Clinical Parameters
2.4. Circulating miR-199b Expression Levels Predict Pathological Response to nCRT and Recurrence in LARC Patients
3. Discussion
4. Materials and Methods
4.1. Cell Cultures and Transfection
4.2. Patient Samples
4.3. Evaluation of Pathological Response
4.4. Wound-Healing Assay
4.5. Transwell Migration Assay
4.6. Colony-Forming Assay
4.7. Cell Viability Assay
4.8. Analysis of Caspase Activation
4.9. Colonosphere Formation Assay
4.10. Western Blot Analysis
4.11. RNA Isolation
4.12. Quantification of miR Expression Levels
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Parameter | No. Cases | No. miR-199b High (%) | No. miR-199 Low (%) | p |
---|---|---|---|---|
MiR-199b | 22 | 17 (77.3) | 5 (22.7) | |
Gender | 22 | 17 | 5 | 0.078 |
Male | 12 | 11 (91.7) | 1 (8.3) | |
Female | 10 | 6 (60) | 4 (40) | |
Age | 22 | 17 | 5 | 0.962 |
<70 | 13 | 10 (76.9) | 3 (23.1) | |
≥70 | 9 | 7 (77.8) | 2 (22.2) | |
Grade pre-CRT 1 | 22 | 17 | 5 | 0.211 |
Low | 14 | 12 (85.7) | 2 (14.3) | |
Moderate-High | 8 | 5 (62.5) | 3 (37.5) | |
Clinical stage pre-CRT | 22 | 17 | 5 | 0.312 |
II | 3 | 3 (100) | 0 (0) | |
III | 19 | 14 (73.7) | 5 (26.3) | |
ECOG 2 | 22 | 17 | 5 | 0.962 |
0 | 13 | 10 (76.9) | 3 (23.1) | |
1 | 9 | 7 (77.8) | 2 (22.2) | |
ypT 3 | 20 | 15 | 5 | 0.002 |
0–2 | 12 | 12 (100) | 0 (0) | |
3–4 | 8 | 3 (37.5) | 5 (62.5) | |
ypN 4 | 20 | 15 | 5 | 0.091 |
0 | 14 | 12 (85.7) | 2 (14.3) | |
1–2 | 6 | 3 (50) | 3 (50) | |
Pathological stage | 20 | 15 | 5 | 0.020 |
yp0-I | 9 | 19 (100) | 0 (0) | |
ypII-III | 11 | 6 (54.5) | 5 (45.5) |
Response to Neoadjuvant CRT 1 | ||||
---|---|---|---|---|
Response | No. Cases | Responders 2 (%) | Non-Responders 3 (%) | p |
MiR-199b expression | 22 | 11 | 11 | 0.011 |
Low | 5 | 0 (0) | 5 (100) | |
High | 17 | 11 (64.7) | 6 (35.3) |
Downstaging | No. Cases | Yes (%) | No (%) | p |
---|---|---|---|---|
MiR-199b expression | 22 | 15 | 7 | 0.009 |
Low | 5 | 1 (20) | 4 (80) | |
High | 17 | 14 (82.4) | 3 (17.6) |
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Santos, A.; Cristóbal, I.; Rubio, J.; Caramés, C.; Luque, M.; Sanz-Alvarez, M.; Morales-Gallego, M.; Madoz-Gúrpide, J.; Rojo, F.; García-Foncillas, J. MicroRNA-199b Deregulation Shows Oncogenic Properties and Promising Clinical Value as Circulating Marker in Locally Advanced Rectal Cancer Patients. Int. J. Mol. Sci. 2022, 23, 2203. https://doi.org/10.3390/ijms23042203
Santos A, Cristóbal I, Rubio J, Caramés C, Luque M, Sanz-Alvarez M, Morales-Gallego M, Madoz-Gúrpide J, Rojo F, García-Foncillas J. MicroRNA-199b Deregulation Shows Oncogenic Properties and Promising Clinical Value as Circulating Marker in Locally Advanced Rectal Cancer Patients. International Journal of Molecular Sciences. 2022; 23(4):2203. https://doi.org/10.3390/ijms23042203
Chicago/Turabian StyleSantos, Andrea, Ion Cristóbal, Jaime Rubio, Cristina Caramés, Melani Luque, Marta Sanz-Alvarez, Miriam Morales-Gallego, Juan Madoz-Gúrpide, Federico Rojo, and Jesús García-Foncillas. 2022. "MicroRNA-199b Deregulation Shows Oncogenic Properties and Promising Clinical Value as Circulating Marker in Locally Advanced Rectal Cancer Patients" International Journal of Molecular Sciences 23, no. 4: 2203. https://doi.org/10.3390/ijms23042203